Salmonella Typhimurium is a major cause of human and animal salmonellosis, impacting global socioeconomic factors. This study examined antibiotic resistance patterns, virulence genes and plasmids in S. Typhimurium isolates from nine South African provinces (1999-2021). Among 180 randomly selected isolates, 129 were confirmed as S. Typhimurium from animal (n = 94), food (n = 16), environment (n = 7) and feed (n = 12) sources using polymerase chain reaction (PCR). Phenotypic resistance was assessed against 13 antibiotics, revealing high resistance levels: 86.8% to ciprofloxacin, 69.0% to ceftriaxone and 65.1% to piperacillin. Multidrug resistance (up to 11 antibiotics) was observed. Genetic analysis identified resistance genes, including blaPSE (32.6%), blaCMY-2 (21.7%), tetA (24.0%), tetB (22.5%), qnrB (21.7%) and qnrA (20.2%). Class 1 integrons were found in 47.4% of isolates. Virulence genes were prevalent, including sopB (95.3%), sspH1 (82.9%), sifA (82.9%), pefA (79.8%), spvC (79.1%), sspH2 (77.5%), srgA (71.3%) and invA (100%). Plasmid analysis detected 2 kb, 8 kb and 90 kb plasmids, with the 90 kb plasmid being most common (71.3%). Enterobacterial Repetitive Intergenic Consensus (ERIC) PCR identified 44 clusters (A-RR), including 6 major clusters.Contribution: These findings highlight the urgent need for enhanced surveillance and intervention strategies to curb antibiotic resistance and virulence in S. Typhimurium populations in South Africa, stressing the importance of monitoring and control measures to address this public health threat.

Brucellosis, a contagious zoonotic bacterial infection affecting livestock and wildlife, is primarily caused by Brucella abortus, globally. However, in South Africa, the true prevalence of bovine brucellosis remains unknown because of a lack of epidemiological data. Therefore, this study used diagnostic data to evaluate and determine the seropositivity of bovine brucellosis based on Rose Bengal test (RBT) screening and confirmation with complement fixation test (CFT) in Limpopo and Free State provinces between 2013 and 2022. The use and limitations of this data were also evaluated based on the bovine brucellosis scheme in South Africa. The study revealed the overall seropositivity of 4.2% (n = 8980/212 440) for bovine brucellosis based on RBT and CFT in series. In Limpopo province, the brucellosis seropositivity was slightly higher at 4.3% (n = 7488/173 011) compared to 3.8% (n = 1492/39 429) in Free State province. Analysis of brucellosis distribution over the study period indicated significant variation (p  0.001) both between and within the provinces. Notably, the highest prevalence in Limpopo occurred during 2013-2017, whereas in Free State, peak prevalence was observed in 2013 and 2016. Challenges preventing an accurate reflection of the brucellosis seropositivity in these provinces for the period include a lack of data on vaccination history and herd status of the samples submitted, as well as the inability to match the CFT results from different laboratories, because some laboratories are only accredited to perform the RBT.Contribution: Insights gained from retrospective studies such as this study can play crucial roles in shaping effective control and preventative measures against bovine brucellosis. Given the challenges in obtaining confirmatory test results, we suggest that brucellosis tests be conducted at a single central laboratory or that the government provides a central database where all laboratories can enter their data. Furthermore, information submitted to the laboratories must make herd and vaccine history compulsory for sample submission to ensure more accurate data.

African swine fever (ASF) is a severe viral haemorrhagic disease affecting domestic pigs, maintained by ASF virus within an ancient sylvatic cycle involving Ornithodoros ticks that inhabit warthog burrows. This study aimed to develop and optimise a multiplex polymerase chain reaction (PCR) assay to differentiate three Ornithodoros species in South Africa - O. moubata, O. phacochoerus and O. waterbergensis, to enhance ASF vector identification and management. The final multiplex PCR effectively distinguished between species with specific amplicon sizes: 180 base pairs (bp) for O. moubata, 640 bp for O. phacochoerus and 1000 bp for O. waterbergensis.Contribution: This refined multiplex PCR method demonstrates proof of concept as a reliable and rapid tool for distinguishing Ornithodoros species and correlating them with their geographical origins. This assay, based on amplification size, provides crucial information about the distribution of these species, which could enhance ASF control efforts.

In a controlled experiment, we compared renal-hepatal, haematological and organ histopathology responses in chickens treated with Crude extracts of Capsicum annuum L. and Carica papaya L. to those treated with synthetic anthelmintics. Twenty-one indigenous seven-week-old chickens were fed on Nuvita® feeds Uganda limited and adlib tap municipal water. They were divided into seven groups of three chickens per group. The treatments were; CPLe (Carica papaya L. ethanol extract), CPLa (Carica papaya L. acetone extract), CAFe (Capsicum annuum L. ethanol extract), CAFa (Capsicum annuum L. acetone extract), levamisole, piperazine and phosphate buffered saline (PBS). CPLe, CPLa, CAFe, CAFa were given at a dose of 0.48 g per bird (1.37 g/kg body weight) as determined from a previous in-vitro experiment. Levamisole hydrochloride was given at 25 mg/kg body weight and piperazine citrate at 100 mg/kg body weight. The control group received 0.2% dimethyl sulfoxide (DMSO) in PBS. The treatments were individually administered orally and repeated on the second day. The birds were bled and sacrificed one week after treatment. Blood was submitted for haematology and harvesting of serum for renal-hepatal function tests. The heart, kidneys and liver were also harvested for histological examinations. CPLe caused significantly raised albumin compared to CAFe (p = 0.02), levamisole caused significantly raised AST compared to CAFe (p = 0.04). All extracts and synthetic anthelmentics increased the number of eosinophils, indicating an increased inflammatory response. CPLa, CAFe, CAFa and levamisole were toxic to the kidneys. All extracts were toxic to the liver except CPLe. Piperazine and levamisole were as well toxic to the liver.Contribution: Plant extracts are not necessarily safer than synthetic anthelmintics and should be used with caution.

Ectoparasites can severely impact wildlife species, both through their feeding behaviour and by facilitating secondary infestations or bacterial infections. Here, we report on two cases of periorbital damage caused by ixodid tick infestations in cheetahs (Acinonyx jubatus), including a rare record of traumatic myiasis in wildlife. Both cheetahs were recently reintroduced to the Marromeu-Coutada Complex in Central Mozambique and had sustained relatively severe injuries. This ostensibly yielded them immunocompromised and thus more vulnerable to tick infestations, particularly when exposed to novel species. These incidents occurred during the peak hot-wet season in which adult tick abundance is likely highest. These cases provide novel insights into the risks of tick infestations for reintroduced cheetahs and the importance of intensive post-release monitoring to allow for timeous veterinary intervention.Contribution: This study falls within the scope of the journal. Ixodid ticks are of veterinary importance for wildlife and domestic animals across Africa because of the associated direct damage and potential spread of tick-borne pathogens. This study investigates two cases of tick infestations in cheetahs recently reintroduced into a tropical environment. The identified risk factors appear to be exposure to novel ectoparasites, injuries, and the hot-wet season. This highlights the need to consider ectoparasite risk when planning wildlife translocations.

The widespread use of parabens has led to their accumulation in aquatic environments. This study examined the effects of methylparaben on rainbow trout, dividing 96 fish into control and treatment groups (1 mg/L, 5 mg/L, and 8 mg/L). Results showed dose-dependent weight loss, altered hepatosomatic indices, increased serum urea, uric acid, and Malondialdehyde (MDA) levels, and decreased Glutathione Peroxidase (GSH-Px) activity. Histopathological analysis revealed liver and kidney abnormalities in treated groups, including hepatocyte degeneration, proliferation in the bile duct, glomerular atrophy, reduced haematopoietic tissue, increased melanomacrophage centres, necrosis and fibrosis.Contribution: These findings highlight methylparaben's toxic effects, emphasising the need for stricter regulations and further research to safeguard aquatic ecosystems and understand its impact on aquatic organisms.

Paratuberculosis (Johne's disease) has devastating outcomes on ruminant health and impacts on national and international trade. The current work assessed the diagnostic value of the VersaTREK automated liquid culture system in isolating Mycobacterium avium subspecies paratuberculosis (MAP) from faecal and intestinal tissue samples from ovine under South African conditions and compared it with the current method of choice, histopathological examination. Intestinal tissue and faecal samples from 111 sheep (including complete set from 104 slaughter sheep from flocks with a history of MAP infection as well as incomplete sample sets from 7 sheep) were analysed using the liquid culture method. One set of tissues was subjected to histopathological examination. Deoxyribonucleic acid (DNA) extracted from culture isolates was subjected to polymerase chain reaction (PCR) amplification using primers that target the IS900 regions of the MAP for species verification. Overall, the VersaTREK automated liquid culture in combination with IS900 PCR showed a comparable level of detection in tissues (12.6%) as histopathology (13.5%), but the detection rate for faecal samples was lower than for tissues (10.8%). A combination of histopathology and faecal culture increased the detection rate from 13.5% (n = 14/104) and 9.6% (n = 10/104), respectively, to 15.4% (n = 16/104).Contribution: Our findings highlight the diagnostic utility of the VersaTREK automated liquid culture system in detecting MAP in ovine samples collected both ante and postmortem. However, an inhibitory effect on the MAP isolation rate observed when the antibiotic cocktail was added to the culture medium warrants further investigation. The outcome of the study is beneficial in guiding the strategic planning of the nationwide control programme.

African swine fever (ASF) is a highly fatal viral haemorrhagic disease affecting pigs. This study uses official surveillance data to investigate the persistence and spread of ASF in wild boar populations in Serbia from 2020 to 2024. A total of 480 ASF cases were analysed, with spatiotemporal clustering conducted via SaTScan™ and proximity analyses performed in the Quantum Geographic Information System. The yearly prevalence of ASF in wild boar rose steadily from 0.66% in 2020 to 1.47% in 2023. Seasonal trends showed a significant increase in cases during winter (68%) and spring (24%). Five distinct clusters were identified, predominantly near the borders with North Macedonia and Bulgaria, suggesting potential cross-border transmission. Proximity to major roads was significantly associated with ASF case distribution (p  0.01), while proximity to water bodies and elevation showed no consistent pattern. The findings highlight critical gaps in current passive surveillance systems, which likely underestimate ASF prevalence. The study underscores the need for enhanced surveillance in remote and wooded areas and proposes strategies to improve ASF monitoring and control in wild boar populations.Contribution: This study highlights the feasibility of cost-effective, non-invasive surveillance methods for ASF detection, offering critical insights for low-income countries where resources for intensive disease monitoring are limited. By demonstrating how environmental and anthropogenic factors drive ASF dynamics, this research provides actionable strategies for improving regional and global ASF control efforts.

Natural treatment options for Microsporum canis dermatophytosis are being explored because of resistance to several antifungal medications. In this study, the potential antifungal effect of bee venom (BV), a natural antimicrobial agent, on M. canis was investigated. The antifungal effects of BV, fluconazole, itraconazole, amphotericin B and terbinafine were evaluated by the macrodilution method at various concentrations by modifying the microdilution method recommended by the European Committee on Antimicrobial Susceptibility Testing. All isolates were observed to be susceptible to terbinafine and fully resistant to fluconazole and amphotericin B. Minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) values of M. canis isolate 2 (Mc2) were determined as 8 µg/mL for itraconazole. The MIC and MFC values of BV were found to be 320 µg/mL for the Mc2 isolate and 640 µg/mL for the Mc6 isolate. The results showed that the isolates obtained from clinical samples in this study were highly resistant to all antifungal agents, except terbinafine. The increase in resistance indicates that antifungal drugs will become insufficient and ineffective over time and natural products such as BV should be evaluated as alternatives.Contribution: Although there are many drugs for the treatment of M. canis, the increase in resistance to antifungal agents reveals the need for the identification and development of new natural agents. Bee venom, which has been shown to have a safe and weak allergenic effect in various studies, can be tested for usability as a local antifungal drug when supported by in vivo studies.

Meat inspection is the routine method used to identify cattle infected with Taenia saginata; however, the sensitivity of this method is low. We investigated the prevalence of T. saginata infection in cattle slaughtered in low throughput abattoirs (LTs) in Gauteng province, South Africa, based on meat inspection and serology. A total of 188 cattle carcasses from three abattoirs underwent meat inspection for the presence of T. saginata cysticerci, while serum was tested for the occurrence of antigens using antigen enzyme-linked immunosorbent assay (Ag-ELISA). Polymerase chain reaction (PCR)-sequencing was performed to confirm T. saginata cysts identified during enhanced inspection and incision of 10 randomly selected hearts. Apparent and true prevalence were calculated, and a logistic regression model was fit to evaluate associations between abattoir, sex, animal origin, age and T. saginata serological status. Out of the 188 carcasses, no positive cases (0%) were identified during routine meat inspection, yet three cysticerci-like lesions were identified from three hearts following additional incisions, of which one was confirmed as T. saginata. Fifty-four of the sampled cattle carcasses tested positive using Ag-ELISA (apparent and true prevalence: 29.0%, 71.8%). Feedlot cattle were less likely (odds ratios [OR]: 0.33, p = 0.043) to have a positive serological test result for T. saginata compared to non-feedlot cattle, and the odds of a positive result differed between abattoirs (p  0.05).Contribution: Our results confirmed the low sensitivity of routine meat inspection in LTs, which may pose a public health risk, and therefore other diagnostic methods need to be included in the surveillance system for T. saginata.

To investigate the existence of bovine viral diarrhoea virus (BVDV), parainfluenza virus 3 (PIV-3) and respiratory syncytial virus (RSV) as well as its coinfections, a total of 420 pneumonic lung tissue samples were collected from slaughterhouses in three different areas. Samples were examined for the three viruses using antigen detection enzyme-linked immunosorbent assay (ELISA) test, and positive results were further confirmed using fluorescent antibody test and polymerase chain reaction. Prevalences detected were 10.47% for BVDV, 11.9% for parainfluenza virus type (PIV-3) and 7.6% for respiratory syncytial virus (RSV). Observed coinfections were bovine viral diarrhoea (BVD) and PIV-3 in 8.1%, BVD and RSV in only two samples (1%). Existence of all three viruses was detected in two samples (1%).Contribution: In this study, infection of the three viruses was confirmed in cattle and existence of its coinfection is documented for the first time.

Plant foliages used as feed additives pose a health risk due to high oxidant concentrations. Oxidants cause oxidative stress and high rate of morbidities and mortalities. Hence, the aim of the study was to validate the methods to quantify gallic acid (GA) and quercetin (Q) as putative antioxidants, and to evaluate antioxidant activities in feed (F), Hyperacanthus amoenus (HA) and Carissa bispinosa (CB) extracts. Extraction was carried out with 62.5% methanol. Method validations for linearity, accuracy and precision were performed on high performance liquid chromatography. Quantitative analysis of GA and Q and testing of 2,2-diphenyl-2-picrylhydrazyl (DPPH) scavenging activities in the extracts were performed. The lowest limit of detection (LOD) and limit of quantitation (LOQ) of 0.011 µg/mL and 0.032 µg/mL were determined in HA, respectively. The methods were accurate and precise as the relative standard deviations (%RSD) were less than 15%. The GA concentrations in CB and HA extracts were statistically significant (p  0.05) and their values were 0.65 ± 0.03 x 106 µg/kg dry weight (DW) (0.13%) and 0.28 ± 0.06 x 106 µg/kg DW (0.002%), respectively. All extracts showed very strong radical scavenging activities with their IC50 values ranging between 5.87 µg/mL and 6.86 µg/mL.Contribution: These accurate, repeatable, precise and reliable methods can be used to provide a valuable basis for GA and Q analysis in various shrub foliages. Though high GA concentrations have potential to act as antioxidants, they may have adverse health and growth performance effects when used as feed additives, while lower Q concentrations may have no effects on livestock.

A cross-sectional study was conducted to assess antibiotic usage, residues, resistance and drivers of their emergence in fish aquaculture in Northern Nigeria between 2019 and 2020. A structured questionnaire was administered to 151 randomly selected farmers. Fish, column and wastewater samples were analysed with enzyme link immunosorbent assay (ELISA) for residue detection and quantification. We performed descriptive and analytic statistical analyses. All selected farmers participated, 78.1% used antibiotics in ponds. Majority (77.1%) of the farmers did not know what antibiotic misuse entailed and 73.3% mentioned that antibiotic misuse and overuse cannot predispose to residues and resistance. The most frequently used antibiotics were tetracyclines (TCs) (99.2%). Significant risk routes for residue and resistance spread were: contaminated fish and its products ingestion, direct contact with fish and fomites contaminated as well as water and aerosols contaminated in the pond environment. Significant social and cultural that influenced residue and resistance development include antibiotics misuse/overuse (Odds Ratio [OR] = 3.8; 95% confidence interval [CI]: 1.62-8.74) as well as poor education and expertise of the farmers (OR = 2.9; 95% CI: 1.24-6.94). The mean TCs residues level in column and wastewater was 123.6 ± 18.2 μg/kg-1. Poor knowledge and attitudes regarding antibiotic usage were identified. Adequate antibiotic stewardship should be promoted through farmers' education. Identified social and cultural factors can be mitigated through the 'One Health' approach.

Anti-nutritive and toxic factors in plants adversely affect animal health and production. Hence, the aims of the study were to (1) carry out qualitative and quantitative analysis of these factors in Terminalia sericea that grows abundantly in Onderstepoort; (2) evaluate an affordable, efficient and reliable method that reduces these factors and (3) evaluate antioxidant properties of the phenolic compounds. Thus, sequential organic and aqueous extractions were carried out on T. sericea leaves and feed pellets. Extracts were subjected to qualitative analysis to confirm the presence of 11 phytochemicals. While quantitative analysis of nine phytochemicals was carried out on spectrophotometry, the phenolic extracts were further tested for antioxidant activity on 2,2-diphenyl-picrylhydrazyl (DPPH) assay where gallic acid (GA), quercetin (Q) and ascorbic acid (AA) were used as standards. The contents of GA, Q and AA in the phenolic extracts were determined on high-performance liquid chromatography (HPLC). Aqueous extraction was an effective method as most phytochemicals were extracted. Nine phytochemicals were qualitatively analysed in T. sericea. The highest contents of 1.97% and 2.37 ± 0.077 mg/g gallic acid equivalent with a significant difference (p  0.05) were determined in oxalates and phenolics, respectively. Furthermore, 83.1% DPPH inhibition (IC50 = 0.0432 µg/mL) was determined relative to the standards (85.27% - 85.96%; IC50 = 0.000009 µg/mL - 0.00042 µg/mL). Subsequently, HPLC results showed that T. sericea contained high concentration of AA (62.5 ± 2.13 µg/mL) to confirm antioxidant properties of the leaves.Contribution: Though T. sericea leaves have antioxidant activities, the alkaloid contents may pose a threat to the livestock. Therefore, farmers around Onderstepoort should protect their livestock from feeding on these leaves.

The COVID-19 pandemic has caused the death of 7.1 million people worldwide as of 7 July 2024. In Nigeria, the first confirmed case was reported on 27 February 2020, subsequently followed by a nationwide spread of SARS-CoV-2 with morbidity and mortality reaching 267 173 and 3155, respectively, as of 7 July 2024. At the beginning of the pandemic, only a few public health laboratories in Nigeria had the capacity for SARS-CoV-2 molecular diagnosis. The National Veterinary Research Institute (NVRI), already experienced in influenza diagnosis, responded to the public health challenge for the diagnosis of COVID-19 samples from humans. The feat was possible through the collective utilisation of NVRI human and material resources, including biosafety facilities, equipment, reagents and consumables donated by international partners and collaborators. Within 6 months of the reported COVID-19 outbreak in Nigeria, over 33 000 samples were processed in NVRI facilities covering five states. Thereafter, many field and laboratory projects were jointly implemented between NVRI and collaborating sectors including the Nigerian Centre for Disease Control (NCDC) and the National Institute for Medical Research (NIMR), which brought together professionals in the health, veterinary, education and socio-sciences. In addition, One Health grants were secured to enhance surveillance for coronavirus and other zoonoses and build capacity in genomics. Bio-surveillance for coronaviruses and other emerging zoonotic pathogens at the human-animal interface was activated and continued with sample collection and analysis in the laboratory for coronaviruses, Lassa fever virus and Mpox. One Health approach has shown that inter-sectoral and multinational collaboration for diagnosis, research and development in animals, and the environment to better understand pathogen spillover events at the human-animal interface is an important global health priority and pandemic preparedness.

Rabies is endemic in Sudan with continuing outbreaks occurring annually, the most common animals affected are dogs, followed by goats and equids. This work focused on equid rabies, to elucidate the current situation of the disease through analysis of reports of equid rabies outbreaks in Sudan during 2010-2022 supported by laboratory confirmation of the disease. During the study period, 66 animals were affected during 35 equid rabies outbreaks. The highest incidences were found in Al Gezira (30.3%), followed by Darfur (24.2%) and Kordofan (15.2%). The highest incidence rate was observed during 2018 (33.3%), followed by 2015 (16.7%). Within seasons, the highest incidence rate was reported during October - December (33.3%), followed by July - September (30.3%). Chi-square analysis revealed a significant correlation between rabid animals and year, season, and state. Wald statistics demonstrated that year and season had a significant association with the disease. Virus antigen was identified (72.2%) in brain tissues using the fluorescent antibody test. Viral nucleic acid was amplified (n = 6) with a reverse transcriptase polymerase chain reaction assay.Contribution: As equids are kept in close contact with humans and other animals in the country, according to the present investigation, equid rabies in Sudan is a potential public health concern, emphasising the importance of implementing effective control measures.

Human rabies transmitted by dogs still kills thousands of people each year worldwide. Dog bites are common in the city of Beni (Democratic Republic of Congo), which shows low rabies vaccination coverage. This study aimed to determine the factors associated with the rabies vaccination status of dogs. A cross-sectional analytical study was conducted in the town of Beni among dog owners, during a household survey selected using a multistage sampling. The information sought concerned the knowledge and characteristics of the dog owners as well as the vaccination status of these dogs. Logistic regression was used to investigate associations between the vaccination status of the dogs and the main independent factors. Rabies vaccination coverage in Beni was 26% (95% confidence interval [CI]: 22% - 30%). The main factors associated with the rabies vaccination status of the dog were primary education level of household head (adjusted odds ratio [aOR]:4.8; 95% CI: 1.2- 19.8); university education level of household head (aOR: 5.9; 95% CI: 1.6-22); perceived rabies severity (aOR: 44. 4; 95% CI: 10.4-188), having more than one dog in the household (aOR: 2.6; 95% CI: 1.6-4.3); age range 7-12 months (aOR: 0.2; 95% CI: 0.1-0.6) and confined dog breeding (aOR: 3.9; 95% CI: 1.1-14.9). The low vaccination coverage in Beni requires mass vaccination campaigns against canine rabies targeting the dog owners with low education levels, those raising more than one dog, with stray dogs or dogs less than 12 months old.

Various zoonotic microorganisms cause reproductive problems such as abortions and stillbirths, leading to economic losses on farms, particularly within livestock. In South Africa, bovine brucellosis is endemic in cattle, and from 2013-2018, outbreaks of Brucella melitensis occurred in sable. Coxiella burnetii, the agent responsible for the zoonotic disease known as Q-fever and/or coxiellosis, also causes reproductive problems and infects multiple domestic animal species worldwide, including humans. However, little is known of this disease in wildlife. With the expansion of the wildlife industry in South Africa, diseases like brucellosis and coxiellosis can significantly impact herd breeding success because of challenges in identifying, managing and treating diseases in wildlife populations. This study investigated samples obtained from aborted sable and roan antelope, initially suspected to be brucellosis, from game farms in South Africa using serology tests and ruminant VetMAX™ polymerase chain reaction (PCR) abortion kit. The presence of C. burnetii was confirmed with PCR in a sable abortion case, while samples from both sable and roan were seropositive for C. burnetii indirect enzyme-linked immunosorbent assay (iELISA). This study represents the initial report of C. burnetii infection in sable and roan antelope in South Africa. Epidemiological investigations are crucial to assess the risk of C. burnetii in sable and roan populations, as well as wildlife and livestock in general, across South Africa. This is important in intensive farming practices, particularly as Q-fever, being a zoonotic disease, poses a particular threat to the health of veterinarians and farm workers as well as domestic animals.Contribution: A report of clinical C. burnetii infection in the wildlife industry contributes towards the limited knowledge of this zoonotic disease in South Africa.

The South African government recognises the value of traditional healing and supports an integrated healthcare system. In the veterinary field, successful integration depends on the perceptions of different animal healthcare practitioners (AHCPs). This study aimed to determine the perceptions of ethnoveterinary medicine among state-employed veterinarians and animal health technicians in the North West province. A mixed model questionnaire and non-probability sampling gathered quantitative and qualitative data from 53 participants. Statistical and thematic analysis processes were applied to quantitative and qualitative data, respectively. Overall, 77.4% of participants were aware of ethnoveterinary medicine practices. Approval of the use of ethnoveterinary medicine by farmers was above-average at a mean of 3.57, standard deviation (s.d.) = 1.016 on a 5-point Likert scale. However, 65.9% doubted the effectiveness of ethnoveterinary medicine, which suggests tolerance but no appreciation of its value. The Chi-square test revealed a significant relationship between district municipality of employment and awareness at χ2 = 17.490, df = 3, p = 0.001. Thematic analysis of comments showed that some participants perceived ethnoveterinary medicine as unscientific and interfering with Western veterinary medicine, while some considered it a good complement to Western veterinary medicine. The study also found that 20.8% of AHCPs use ethnoveterinary medicine to treat certain diseases. In conclusion, the lack of appreciation for ethnoveterinary medicine among AHCPs may hinder government-proposed integration, emphasising the need for programmes to improve perceptions among these stakeholders.Contribution: This is first such study to report on perceptions of ethnoveterinary medicine among AHCPs, and assess their readiness for an integrated animal health system.

Wild animals, sharing pathogens with domestic animals, play a crucial role in the epidemiology of infectious diseases. Sampling from wild animals poses significant challenges, yet it is vital for inclusion in disease surveillance and monitoring programmes. Often, mass surveillance involves serological screenings using enzyme-linked immunosorbent assay (ELISA) tests, typically validated only for domestic animals. This study assessed the diagnostic specificity of commercially available ELISA tests on 342 wild ruminant serum samples and 100 from wild boars. We evaluated three tests for foot-and-mouth disease: two for Peste des petits ruminants, two for Rift Valley fever and one for Capripox virus. Diagnostic specificity was calculated using the formula True Negative/(False Positive + True Negative). Cohen's kappa coefficient measured agreement between tests. Results showed high specificity and agreement across all tests. Specificity for foot-and-mouth disease (FMD) ranged from 93.89% for Prionics to 100% for IDEXX, with IDvet showing 99.6%. The highest agreement was between FMD IDvet and IDEXX at 97.1%. Rift Valley fever (RVF) tests, Ingezim and IDvet, achieved specificities of 100% and 98.83%, respectively. The optimal specificity was attained by retesting single reactors and inactivating the complement.Contribution: Commercially available ELISA kits are specific for foot-and-mouth disease and similar transboundary animal diseases and can be used for highly specific wild animal testing.